A number of studies have shown that various oligonucleotides that are complementary to DNA or RNA (including mRNA) can hybridize to the genetic material and inhibit transcription, replication or translation.
It has been shown by Smith et al., Proc. Natl. Acad. Sci. U.S.A., 83: 2787-91 (1986) that an eight residue oligomer complementary to the acceptor splice junction of Herpes Simplex Virus-1 (HSV-1) immediate early mRNAs 4 and 5 inhibits viral replication with little, if any, deleterious effect on host-cell macromolecular metabolism and growth rate. It has also been shown that sequences that are complementary to regions immediately adjacent to the tRNA Lys primer binding site in human T-cell lymphotropic virus type III (HTLV-III) inhibited replication of the virus [Zamecnik et al., Proc. Natl. Acad. Sci. U.S.A., 83: 4143-6 (1986)]. Similar inhibition was seen in the same study when the oligomers were complementary to a splice donor site or a splice acceptor site. The seven oligomers of the Zamecnik et al. study varied in size from 12 to 26 bases. In another study, a 13-mer (an oligomer containing 13 repeating units) that is complementary to the reiterated terminal sequence of Rous sarcoma virus (RSV) 70s RNA inhibits the production of RSV with the proposed mechanism of inhibition being either in the circularization step of the proviral DNA intermediate and/or in the initiation of translation [Stephenson and Zamecnik, Proc. Natl. Acad. Sci. U.S.A., 75: 285-4 (1978)]. U.S. Pat. No. 4,757,055 discloses certain oligonucleoside alkyl and aryl phosphonates that are complementary to base sequences "foreign" to mammalian cells and that hinder the replication and/or translation of such foreign base sequences. The foreign sequences relating to HSV relate to the acceptor splice junction (column 23, lines 12-14).
European Patent Application No. 263740 discloses RNA and DNA sequences that are complementary to the ribosome binding site of mRNAs, and to sites that are critical to viral RNA polymerase binding (page 4, lines 57 through 64). The various sequences disclosed are postulated to have antiviral effects.
In non-viral systems, inhibition of translation has been postulated to occur in various systems. Expression of the genes coding for the major outer membrane proteins in E. coli is inhibited by a 174-base mRNA (referred to as mRNA--interfering complementary RNA or micRNA) that is transcribed by the E. coli and that contains a sequence that is complementary to the 5' end region of the membrane protein mRNAs [Mizuno et al., Proc. Natl. Acad. Sci. U.S.A., 81: 1966-70 (1984)]. In frog oocytes, microinjection of RNA complementary to globin mRNA (anti-sense globin RNA) formed a hybrid with globin mRNA and selectively prevented translation [Melton, Proc. Natl. Acad. Sci. U.S.A., 82: 144-8 (1985)]. In rabbits, inhibition of rabbit globin mRNA translation by oligonucleotides complementary to the 5' end region has been observed [Blake et al., Biochemistry, 24: 6132-45 (1985)].
Non-ionic versions of complementary oligodeoxyribonucleoside methylphosphonates were shown to inhibit translation of dihydrofolate reductase (DHFR) to a lesser extent than the corresponding oligodeoxyribonucleotides [Maher and Dolnick, Nucleic Acids Res., 16: 3341-58 (1988)].
In HSV (herpes simplex virus) one of the factors affecting selective initiation of alpha phase gene transcription is a transactivating virion component, the Vmw65 protein [J. Mol. Biol., 180:1-19 (1984)]. Some transactivating virion components, including Vmw65, may be produced from the viral genetic material and stored in new virus particles. After invasion of a new host cell by these virus particles, the transactivating components interact with certain regions of the viral DNA sequences and stimulate transcription of the adjacent downstream genes, which code for proteins needed by the virus for replication. Optimal activation by Vmw65 requires specific DNA structural elements, including an activation-response DNA sequence recognized by the Vmw65 protein. This activation-response DNA sequence is situated usually near the alpha phase promoter so that recognition of the activation-response DNA sequence by the transactivating Vmw65 protein stimulates transcription of the gene located downstream from the alpha phase promoter. A number of proteins found in Herpes Simplex Virus have similar transactivating potential when their corresponding activation-response DNA sequences are located upstream from target genes. These proteins include the proteins ICP4, ICP0 and ICP27, which are described in Everett, J. Gen. Virol. 67:2507-13(1986); and Rice and Knipe, J. Virol. 62(10):3814-23 (1988). Similar transactivating mechanisms exist, for example, in the Pseudorabies virus (PRV-IE gene product), Adenovirus (Ad-EIA) and Human cytomegalovirus (HCMV-MIE protein).
Interference with or disruption of the viral transactivating mechanism could ultimately result in the inhibition of viral replication. To test this theory I have invented a method whereby the normal translation of certain target transactivating proteins is prevented. The results of the practice of this method, which are reported herein, demonstrate that inhibition of viral replication has been accomplished.